Microglia show differential transcriptomic response to Aβ peptide aggregates ex vivo and in vivo

McFarland, Karen N.; Ceballos, Carolina Wiesner; Rosario, Awilda M.; Ladd, Thomas B.; Moore, Brenda D.; Golde, Griffin; Wang, Xue; Allen, Mariet; Ertekin-Taner, Nilüfer; Funk, Cory C.; Robinson, Max; Baloni, Priyanka; Rappaport, Noa; Chakrabarty, Paramita; Golde, Todd E.

doi:10.26508/lsa.202101108

Cited by 21 publications

(21 citation statements)

References 75 publications

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“…Microglia in vitro and in the brain also have different responses to Aβ. Murine primary microglia display rapid and substantial transcriptional changes in response to Aβ, while in vivo microglia do not display the same gene expression changes, suggesting that primary microglia poorly recapitulate in vivo conditions ( McFarland et al, 2021 ). Nevertheless, studies have shown that culturing microglia together with neurons can mitigate this problem.…”

Section: Introductionmentioning

confidence: 99%

A Neuron, Microglia, and Astrocyte Triple Co-culture Model to Study Alzheimer’s Disease

Luchena

Zuazo-Ibarra

Valero

et al. 2022

Front. Aging Neurosci.

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Glial cells are essential to understand Alzheimer’s disease (AD) progression, given their role in neuroinflammation and neurodegeneration. There is a need for reliable and easy to manipulate models that allow studying the mechanisms behind neuron and glia communication. Currently available models such as co-cultures require complex methodologies and/or might not be affordable for all laboratories. With this in mind, we aimed to establish a straightforward in vitro setting with neurons and glial cells to study AD. We generated and optimized a 2D triple co-culture model with murine astrocytes, neurons and microglia, based on sequential seeding of each cell type. Immunofluorescence, western blot and ELISA techniques were used to characterize the effects of oligomeric Aβ (oAβ) in this model. We found that, in the triple co-culture, microglia increased the expression of anti-inflammatory marker Arginase I, and reduced pro-inflammatory iNOS and IL-1β, compared with microglia alone. Astrocytes reduced expression of pro-inflammatory A1 markers AMIGO2 and C3, and displayed a ramified morphology resembling physiological conditions. Anti-inflammatory marker TGF-β1 was also increased in the triple co-culture. Lastly, neurons increased post-synaptic markers, and developed more and longer branches than in individual primary cultures. Addition of oAβ in the triple co-culture reduced synaptic markers and increased CD11b in microglia, which are hallmarks of AD. Consequently, we developed a straightforward and reproducible triple co-cultured model, where cells resemble physiological conditions better than in individual primary cultures: microglia are less inflammatory, astrocytes are less reactive and neurons display a more mature morphology. Moreover, we are able to recapitulate Aβ-induced synaptic loss and CD11b increase. This model emerges as a powerful tool to study neurodegeneration and neuroinflammation in the context of AD and other neurodegenerative diseases.

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Section: Introductionmentioning

confidence: 99%

A Neuron, Microglia, and Astrocyte Triple Co-culture Model to Study Alzheimer’s Disease

Luchena

Zuazo-Ibarra

Valero

et al. 2022

Front. Aging Neurosci.

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“…In addition, the cellular and anatomical complexity of in vivo models may limit further mechanistic insight and cell type-specific intervention in the communication between neurons, astrocytes and microglia. On the other hand, 2D in vitro monocultures lack the in vivo complexity to accurately model the physiological and pathological responses of neural cells in general, and microglia in particular [ 67 , 69 ] similar to astrocytes, as mentioned above. For ß-amyloid formation, a 3D neuron/astrocyte/microglia tri-culture has been developed using immortalized microglia [ 28 ], but no 3D tri-culture of tau aggregation exists yet.…”

Section: Discussionmentioning

confidence: 99%

A 3D human co-culture to model neuron-astrocyte interactions in tauopathies

et al. 2023

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Background Intraneuronal tau aggregation is the major pathological hallmark of neurodegenerative tauopathies. It is now generally acknowledged that tau aggregation also affects astrocytes in a cell non-autonomous manner. However, mechanisms involved are unclear, partly because of the lack of models that reflect the situation in the human tauopathy brain. To accurately model neuron-astrocyte interaction in tauopathies, there is a need for a model that contains both human neurons and human astrocytes, intraneuronal tau pathology and mimics the three-dimensional architecture of the brain. Results Here we established a novel 100–200 µm thick 3D human neuron/astrocyte co-culture model of tau pathology, comprising homogenous populations of hiPSC-derived neurons and primary human astrocytes in microwell format. Using confocal, electron and live microscopy, we validate the procedures by showing that neurons in the 3D co-culture form pre- and postsynapses and display spontaneous calcium transients within 4 weeks. Astrocytes in the 3D co-culture display bipolar and stellate morphologies with extensive processes that ensheath neuronal somas, spatially align with axons and dendrites and can be found perisynaptically. The complex morphology of astrocytes and the interaction with neurons in the 3D co-culture mirrors that in the human brain, indicating the model’s potential to study physiological and pathological neuron-astrocyte interaction in vitro. Finally, we successfully implemented a methodology to introduce seed-independent intraneuronal tau aggregation in the 3D co-culture, enabling study of neuron-astrocyte interaction in early tau pathogenesis. Conclusions Altogether, these data provide proof-of-concept for the utility of this rapid, miniaturized, and standardized 3D model for cell type-specific manipulations, such as the intraneuronal pathology that is associated with neurodegenerative disorders.

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“…The significance of the association ( p -value) was reported following Tukey’s correction. To calculate the cell-type-specific DEGs and neurodegeneration-associated glial gene expression signatures, the geometric means of a set of genes characterizing each profile were used, as described before [ 69 ]. Briefly, we used a transcriptomic database which compiled the expression from RNA sequencing from different cell types (neurons, astrocytes, microglia, newly formed oligodendrocytes, endothelial cells and pericytes) in the brain [ 70 ] to generate lists of genes whose expression was enriched in each representative cell type ( Table S2 ).…”

Section: Methodsmentioning

confidence: 99%

MhcII Regulates Transmission of α-Synuclein-Seeded Pathology in Mice

Cruz

Moon

et al. 2022

IJMS

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MHCII molecules, expressed by professional antigen-presenting cells (APCs) such as T cells and B cells, are hypothesized to play a key role in the response of cellular immunity to α-synuclein (α-syn). However, the role of cellular immunity in the neuroanatomic transmission of α-syn pre-formed fibrillar (PFF) seeds is undetermined. To illuminate whether cellular immunity influences the transmission of α-syn seeds from the periphery into the CNS, we injected preformed α-syn PFFs in the hindlimb of the Line M83 transgenic mouse model of synucleinopathy lacking MhcII. We showed that a complete deficiency in MhcII accelerated the appearance of seeded α-syn pathology and shortened the lifespan of the PFF-seeded M83 mice. To characterize whether B-cell and T-cell inherent MhcII function underlies this accelerated response to PFF seeding, we next injected α-syn PFFs in Rag1−/− mice which completely lacked these mature lymphocytes. There was no alteration in the lifespan or burden of endstage α-syn pathology in the PFF-seeded, Rag1-deficient M83+/− mice. Together, these results suggested that MhcII function on immune cells other than these classical APCs is potentially involved in the propagation of α-syn in this model of experimental synucleinopathy. We focused on microglia next, finding that while microglial burden was significantly upregulated in PFF-seeded, MhcII-deficient mice relative to controls, the microglial activation marker Cd68 was reduced in these mice, suggesting that these microglia were not responsive. Additional analysis of the CNS showed the early appearance of the neurotoxic astrocyte A1 signature and the induction of the Ifnγ-inducible anti-viral response mediated by MhcI in the MhcII-deficient, PFF-seeded mice. Overall, our data suggest that the loss of MhcII function leads to a dysfunctional response in non-classical APCs and that this response could potentially play a role in determining PFF-induced pathology. Collectively, our results identify the critical role of MhcII function in synucleinopathies induced by α-syn prion seeds.

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Microglia show differential transcriptomic response to Aβ peptide aggregates ex vivo and in vivo

Cited by 21 publications

References 75 publications

A Neuron, Microglia, and Astrocyte Triple Co-culture Model to Study Alzheimer’s Disease

A Neuron, Microglia, and Astrocyte Triple Co-culture Model to Study Alzheimer’s Disease

A 3D human co-culture to model neuron-astrocyte interactions in tauopathies

MhcII Regulates Transmission of α-Synuclein-Seeded Pathology in Mice

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